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Quantitation of Digoxin in Human Plasma Using Negative Ion Electrospray Ionization LC/MS

D. Garteiz, S. Din, and R. Morrow
TexMS Analytical Services, Houston, TX

Introduction

A Varian 1200L LC/MS/MS system was used for quantitative measurement down to 15 picograms of the cardiac glycoside, Digoxin, in human plasma using Electrospray Ionization (ESI)-LC/MS in the negative ion detection mode. Since cardiac glycosides are very thermally labile, ESI is preferred over a hot vaporization process like APCI.

Digoxin is classified as a digitalis drug and is used to treat congestive heart failure and heart rhythm irregularities. Digoxins mode of action is to strengthen the force of the heart contractions, and thus help to regulate the rate and rhythm of the heartbeat. The therapeutic level of Digoxin in human serum is 0.5 - 2.0 ng/mL with toxicity starting at 4.0 ng/mL. Because of Digoxins high potency, doses are administered from 0.125 mg to a maximum of 0.5 mg, once a day. Periodic plasma level measurements are critical for establishing the proper dose administration. The onset of action from administration is from 30 minutes to two hours and when combined with the low dosing regime, plasma levels are expected to be in the low picogram to low nanogram levels. These low picogram levels are easily detected with a Varian 1200L LC/MS system in ESI negative mode.

Instrumentation

Varian 1200L LC/MS equipped with ESI source Materials and Reagents

Digitoxin ordered from Sigma-Aldrich Corp.,Catalog Number D5878

Digoxin ordered from Sigma-Aldrich Corp.,Catalog Number D6003

Ammonium trifluoroacetate (ATFA) Sigma-Aldrich Corp.,Catalog Number D23873-2

All other chemicals are reagent grade or HPLC grade

Sample Preparation

A 1.0 mL of human plasma containing Digoxin is spiked with Digitoxin as the internal standard (Figure 1)
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